• biophysics
• cell biology
• computational biology and bioinformatics
• mathematics and computing

• Spheroids, Cellular/metabolism
• Oxygen/metabolism
• Diffusion
• Humans
• Cell Survival
• Tissue Engineering/methods
• Models, Biological
• Cell Culture Techniques, Three Dimensional/methods

• "Spheroid Revolution" Skoltech Translational Research and Innovation Program
• 075-15-2019-1661 Ministry of Science and Higher Education of the Russian Federation

• angiogenesis
• co-culture
• collagen
• fibrin
• human platelet lysate
• hydrogels

• DFG-NE650/9-1 DFG
• OC1-3 IZKF
• 495328185 DFG

• cell‐instructive hydrogels
• design of experiments
• high‐throughput screening
• in vitro tissue and disease models
• multiple linear regression
• vasculogenesis

• Humans
• Human Umbilical Vein Endothelial Cells/metabolism
• Neovascularization, Physiologic/drug effects
• Hydrogels/chemistry
• Coculture Techniques/methods
• High-Throughput Screening Assays/methods
• Mesenchymal Stem Cells/cytology
• Mesenchymal Stem Cells/metabolism
• Pericytes/cytology
• Pericytes/metabolism
• Pericytes/drug effects
• Cell Culture Techniques/methods
• Cell Culture Techniques/instrumentation
• Endothelial Cells/cytology
• Endothelial Cells/metabolism

• Deutsche Forschungsgemeinschaft
• F. Hoffmann-La Roche

• antiangiogenic therapies
• bevacizumab
• micro-capillaries
• recovery

• Humans
• Human Umbilical Vein Endothelial Cells
• Mesenchymal Stem Cells
• Coculture Techniques
• Hydrogels/pharmacology
• Neoplasms
• Neovascularization, Physiologic

• 310030E_202429 Swiss National Science Foundation
• 310030_169808 Swiss National Science Foundation
• 205321_204318 Swiss National Science Foundation
• CRSII5_186271 Swiss National Science Foundation

• cancer
• cell biology
• oncology

• angiogenesis
• biomaterials
• gene signature
• hydrogel
• molecular signaling
• regenerative medicine

• Hydrogels/chemistry
• Extracellular Matrix/metabolism
• Tissue Engineering
• Biocompatible Materials/chemistry
• Cell Adhesion

• Alzheimer’s disease
• aging
• angiogenesis
• cellular senescence
• extracellular vesicles
• oxidative stress

• КП-06-Н31/16; КП-06-М61/5 National Science Fund of Bulgaria

• 3D tumor simulation
• Agent-based model
• Angiogenesis
• Combination therapy
• Hybrid model
• Vascular tumor growth model

• R01 CA240589 NCI NIH HHS

• cancer
• engineered vasculature
• microfluidics
• organ-on-a-chip
• regenerative medicine

• R34 NS126032 NINDS NIH HHS
• R01 GM126571 NIGMS NIH HHS
• R01AR073135 NIH HHS
• R01 AR074234 NIAMS NIH HHS
• R01AR074234 NIH HHS
• R34NS126032 NIH HHS
• R01 AR073135 NIAMS NIH HHS
• UH3TR003148 NIH HHS
• R01 CA257558 NCI NIH HHS
• R01GM126571 NIH HHS
• UH3 TR003148 NCATS NIH HHS

• 3D tumor models
• cervical cancer
• immunotherapy
• patient-derived organoids (PDOs)
• pre-clinical drug tests
• tumor microenvironment (TME)
• tumor spheroids

• n.a. Comprehensive Cancer Center Niedersachsen
• n.a. Bruno und Helene Jöster Foundation
• n.a. Deutsche Forschungsgemeinschaft
• n.a. Federal Ministry of Education and Research
• MWK: ZN3440 REBIRTH Center for Translational Regenerative Medicine

• Organoid
• Precision medicine
• Regenerative medicine
• Stem cells

• 3D in vitro cancer model
• Breast cancer spheroids
• Calreticulin
• Cancer targeting
• Carboxymethyl cellulose
• Colorectal cancer spheroids
• Gelatin
• Gold nanoclusters

• composite hydrogel
• degradation
• enzymatic crosslinking
• hyaluronic acid
• in-vitro angiogenesis model

Glioblastoma multiforme (GBM) is a rare, yet devastating, primary brain tumor in adults. Current treatments remain generally ineffective and GBM almost invariably recurs, resulting in median survival of 15 months. This high malignancy sources notably from the resilience and invasive capabilities of tumor cells. Within GBM, exists a population of self-sustaining transformed cells with stem-like properties (GSCs), which are thought to be responsible for tumor initiation, growth, and invasion, as well as recurrence. In the tumor microenvironment, GSCs might be found in the vicinity of brain endothelial cells, which provide a protective habitat. Likewise, these resistant, quiescent GSCs may accumulate in hypoxic zones, away from the perivascular niche, or travel towards the healthy brain parenchyma, by eminently co-opting neuro-vascular tracks. Herein, we established an ex vivo model to explore GSC invasive behavior. We found that patient-derived cells massively invade the collagen matrix. In addition, we described that the glycoprotein Neuropilin-1 (NRP1) contributes to GSC spreading and invasion. Indeed, both RNA interference-mediated silencing and CRISPR-mediated gene editing deletion of NRP1 strongly impaired the 3D invasive properties of patient-derived GSCs and their close localization to the brain blood vessels. Of note, other typical features of GSCs, such as expansion and self-renewal were maintained. From a mechanistic standpoint, this biological effect might rely on the expression of the β3 subunit integrin cell-extracellular matrix adhesive receptor. Our data, therefore, propose a reliable approach to explore invasive properties of patient glioma cells ex vivo and identify NRP1 as a mediator in this malignant process.

• Ligue Contre le Cancer
• Fondation ARC pour la Recherche sur le Cancer
• Institut National Du Cancer
• Agence Nationale de la Recherche

Vascular-disrupting agents are an interesting class of anticancer compounds because of their combined mode of action in preventing new blood vessel formation and disruption of already existing vasculature in the immediate microenvironment of solid tumors. The validation of vascular disruption properties of these drugs in vitro is rarely addressed due to the lack of proper in vitro angiogenesis models comprising mature and long-lived vascular-like networks. We herein report an indirect coculture model of human umbilical vein endothelial cells (HUVECs) and human dermal fibroblasts (HDFs) to form three-dimensional profuse vascular-like networks. HUVECs embedded and sandwiched in the collagen scaffold were cocultured with HDFs located outside the scaffold. The indirect coculture approach with the vascular endothelial growth factor (VEGF) producing HDFs triggered the formation of progressively maturing lumenized vascular-like networks of endothelial cells within less than 7 days, which have proven to be viably maintained in culture beyond day 21. Molecular weight-dependent Texas red-dextran permeability studies indicated high vascular barrier function of the generated networks. Their longevity allowed us to study the dose-dependent response upon treatment with the three known antiangiogenic and/or vascular disrupting agents brivanib, combretastatin A4 phosphate (CA4P), and 6´-sialylgalactose (SG) via semi-quantitative brightfield and qualitative confocal laser scanning microscopic (CLSM) image analysis. Compared to the reported data on in vivo efficacy of these drugs in terms of antiangiogenic and vascular disrupting effects, we observed similar trends with our 3D model, which are not reflected in conventional in vitro angiogenesis assays. High-vascular disruption under continuous treatment of the matured vascular-like network was observed at concentrations ≥3.5 ng·ml⁻¹ for CA4P and ≥300 nM for brivanib. In contrast, SG failed to induce any significant vascular disruption in vitro. This advanced model of a 3D vascular-like network allows for testing single and combinational antiangiogenic and vascular disrupting effects with optimized dosing and may thus bridge the gap between the in vitro and in vivo experiments in validating hits from high-throughput screening. Moreover, the physiological 3D environment mimicking in vitro assay is not only highly relevant to in vivo studies linked to cancer but also to the field of tissue regeneration.

• Bundesministerium für Bildung und Forschung

• IL‑8
• Notch
• aldehyde dehydrogenase 1A1
• angiogenesis
• melanoma
• multicellular 3D system
• stemness

• Extracellular Matrix/metabolism
• Humans
• Kallikreins/metabolism
• Neoplasms
• Signal Transduction
• Tumor Microenvironment

Tumors exist in a complex, three-dimensional environment which helps them to survive, grow, metastasize, and resist drug treatment. Simple, reproducible, in vitro models of this environment are necessary in order to better understand tumor behavior. Naturally derived polymers are great 3D cell culture substrates, but they often lack the tunability and batch-to-batch consistency which can be found in synthetic polymer systems. In this review, we describe the current state of and future directions for tumor microenvironment models in synthetic hydrogels.

The tumor microenvironment (TME) plays a determining role in everything from disease progression to drug resistance. As such, in vitro models which can recapitulate the cell–cell and cell–matrix interactions that occur in situ are key to the investigation of tumor behavior and selecting effective therapeutic drugs. While naturally derived matrices can retain the dimensionality of the native TME, they lack tunability and batch-to-batch consistency. As such, many synthetic polymer systems have been employed to create physiologically relevant TME cultures. In this review, we discussed the common semi-synthetic and synthetic polymers used as hydrogel matrices for tumor models. We reviewed studies in synthetic hydrogels which investigated tumor cell interactions with vasculature and immune cells. Finally, we reviewed the utility of these models as chemotherapeutic drug-screening platforms, as well as the future directions of the field.

• hydrogels
• tumor immunity models
• tumor microenvironment
• tumor vasculature models

• Breast circulating tumor
• Migration
• Nuclear lamina protein A/C
• Shear stress
• Yes-associated protein

• Angiogenesis
• Gastric cancer
• Hypoxia
• Prognosis
• Tumor microvessels
• Vascular endothelial growth factor

• Biomarkers, Tumor
• Humans
• Microvessels
• Neovascularization, Pathologic
• Prognosis
• Stomach Neoplasms

In this focused review, we address the role of the kallikrein-related peptidase 3 (KLK3), also known as prostate-specific antigen (PSA), in the regulation of angiogenesis. Early studies suggest that KLK3 is able to inhibit angiogenic processes, which is most likely dependent on its proteolytic activity. However, more recent evidence suggests that KLK3 may also have an opposite role, mediated by the ability of KLK3 to activate the (lymph)angiogenic vascular endothelial growth factors VEGF-C and VEGF-D, further discussed in the review.

• KLK3
• PSA
• VEGF-C
• VEGF-D
• angiogenesis
• cancer
• proteolysis

• Gene Expression Regulation, Neoplastic
• Humans
• Kallikreins/metabolism
• Male
• Neovascularization, Pathologic/metabolism
• Prostate-Specific Antigen/metabolism
• Prostatic Neoplasms/genetics
• Prostatic Neoplasms/metabolism
• Prostatic Neoplasms/physiopathology
• Vascular Endothelial Growth Factor C
• Vascular Endothelial Growth Factor D

• Bioprinting
• CAR
• Cancer
• Immunotherapy
• Organoid
• Spheroid
• TME
• Tumor-on-chip

• clinical trials
• in vitro models
• in vivo models
• melanoma
• preclinical research

• Animals
• Clinical Trials as Topic
• Drug Approval/legislation & jurisprudence
• Drug Discovery/legislation & jurisprudence
• Humans
• Marketing
• Melanoma/drug therapy
• Skin Neoplasms/drug therapy

• UIDP/04138/20 Fundação para a Ciência e Tecnologia, FCT
• PTDC/MED-QUI/31721/2017 Fundação para a Ciência e Tecnologia, FCT
• SFRH/BD/117586/2016 Fundação para a Ciência e Tecnologia, FCT
• UIDB/04138/2020 Fundação para a Ciência e Tecnologia, FCT

• Cancer-associated fibroblasts
• Hydrogel
• In vitro models
• Microfluidics
• Stromal heterogeneity
• Tumor microenvironment
• Tumor-associated macrophages
• Tumoral niches

• Animals
• Extracellular Matrix/pathology
• Humans
• Neoplasms/pathology
• Stromal Cells/pathology
• Tissue Engineering/methods
• Tumor Microenvironment

• 3D model
• tumour
• tumour microenvironment

Tumor stroma is known to significantly influence cancer initiation and progression. In the last decade, 3D cell cultures have shown potential in modeling the tumor microenvironment. This review summarizes the main features of current 3D models, shedding light on their importance in the study of cancer biology and treatment.

It is now well established that the tumor microenvironment plays a key role in determining cancer growth, metastasis and drug resistance. Thus, it is fundamental to understand how cancer cells interact and communicate with their stroma and how this crosstalk regulates disease initiation and progression. In this setting, 3D cell cultures have gained a lot of interest in the last two decades, due to their ability to better recapitulate the complexity of tumor microenvironment and therefore to bridge the gap between 2D monolayers and animal models. Herein, we present an overview of the 3D systems commonly used for studying tumor–stroma interactions, with a focus on recent advances in cancer modeling and drug discovery and testing.

• 3D
• angiogenesis
• cancer therapy
• cell culture
• extracellular matrix
• fibroblasts
• immune system
• in vitro
• stroma
• tumor microenvironment

• 3D bioprinting
• biomaterials
• extracellular matrix (ECM)
• high-content screening
• nanoparticles (NPs)
• organoid
• spheroid
• tumor microenvironment (TME)

Epithelial ovarian cancer (EOC) is a highly heterogeneous disease with a high death rate mainly due to the metastatic spread. The expression of MDM4, a well-known p53-inhibitor, is positively associated with chemotherapy response and overall survival (OS) in EOC. However, the basis of this association remains elusive. We show that in vivo MDM4 reduces intraperitoneal dissemination of EOC cells, independently of p53 and an immune-competent background. By 2D and 3D assays, MDM4 impairs the early steps of the metastatic process. A 3D-bioprinting system, ad hoc developed by co-culturing EOC spheroids and endothelial cells, showed reduced dissemination and intravasation into vessel-like structures of MDM4-expressing cells. Consistent with these data, high MDM4 levels protect mice from ovarian cancer-related death and, importantly, correlate with increased 15 y OS probability in large data set analysis of 1656 patients. Proteomic analysis of EOC 3D-spheroids revealed decreased protein synthesis and mTOR signaling, upon MDM4 expression. Accordingly, MDM4 does not further inhibit cell migration when its activity towards mTOR is blocked by genetic or pharmacological approaches. Importantly, high levels of MDM4 reduced the efficacy of mTOR inhibitors in constraining cell migration. Overall, these data demonstrate that MDM4 impairs EOC metastatic process by inhibiting mTOR activity and suggest the usefulness of MDM4 assessment for the tailored application of mTOR-targeted therapy.

• antimicrobial functions
• cell-cell interactions
• in vitro models
• infection
• inflammation
• innate immunity
• microfluidics
• neutrophil

• Animals
• Aspergillosis/immunology
• Aspergillosis/microbiology
• Aspergillus fumigatus/immunology
• Aspergillus fumigatus/physiology
• Cell Communication/immunology
• Humans
• Immunity, Innate/immunology
• Inflammation/immunology
• Inflammation/microbiology
• Neutrophils/cytology
• Neutrophils/immunology
• Neutrophils/microbiology
• Pseudomonas Infections/immunology
• Pseudomonas Infections/microbiology
• Pseudomonas aeruginosa/immunology
• Pseudomonas aeruginosa/physiology

• T32 HL007899 NHLBI NIH HHS

The stromal microenvironment of breast tumors, namely the vasculature, has a key role in tumor development and metastatic spread. Tumor angiogenesis is a coordinated process, requiring the cooperation of cancer cells, stromal cells, such as fibroblasts and endothelial cells, secreted factors and the extracellular matrix (ECM). In vitro models capable of capturing such complex environment are still scarce, but are pivotal to improve success rates in drug development and screening. To address this challenge, we developed a hybrid alginate-based 3D system, combining hydrogel-embedded mammary epithelial cells (parenchymal compartment) with a porous scaffold co-seeded with fibroblasts and endothelial cells (vascularized stromal compartment). For the stromal compartment, we used porous alginate scaffolds produced by freeze-drying with particle leaching, a simple, low-cost and non-toxic approach that provided storable ready-to-use scaffolds fitting the wells of standard 96-well plates. Co-seeded endothelial cells and fibroblasts were able to adhere to the surface, spread and organize into tubular-like structures. For the parenchymal compartment, a designed alginate gel precursor solution load with mammary epithelial cells was added to the pores of pre-vascularized scaffolds, forming a hydrogel in situ by ionic crosslinking. The 3D hybrid system supports epithelial morphogenesis in organoids/tumoroids and endothelial tubulogenesis, allowing heterotypic cell-cell and cell-ECM interactions, while presenting excellent experimental tractability for whole-mount confocal microscopy, histology and mild cell recovery for down-stream analysis. It thus provides a unique 3D in vitro platform to dissect epithelial-stromal interactions and tumor angiogenesis, which may assist in the development of selective and more effective anticancer therapies.

• alginate
• angiogenesis
• hydrogel
• organoid
• outgrowth endothelial cells
• tissue engineering
• vascularized stroma